Leptin mutation and mycobacterial infection lead non-synergistically to a similar metabolic syndrome

The leptin signaling pathway plays an important role as a key regulator of glucose homeostasis, metabolism control and systemic inflammatory responses. However, the metabolic effects of leptin on infectious diseases, for example tuberculosis, is still little known. For transcriptome studies of zebrafish larvae, deep RNA sequencing was used. Mycobacteria induced a very distinct transcriptome signature in the lepbibl54 mutant zebrafish compared to wild type sibling control larvae. Zebrafish lepb+ and lepbibl54 embryos were collected from 6-month-old wild type and lepbibl54 mutant parents, respectively. The embryos were injected into yolk with M. marinum strain M labelled with mWasabi plasmid pTEC15 vector22 or mock injected with 2% polyvinylpyrrolidone 40 (pvp) at 4 to 6 hour hours post fertilization (hpf). M. marinum preparation were followed by the protocol of a previous study (Benard et al., 2012). Microinjection of zebrafish embryos was performed using an automatic microinjection system (Life Science Methods) described previously (Spaink et al., 2013). Zebrafish larvae at 5 days post fertilization (dpf) were collected and stored at -80℃ for RNA isolation.